_r IThis University of"Wisconsin Bioengineering Research Partnership proposes to develop technology capable of rapid, linexpcnsivc, and robust production of long dsDNA molecules (genes) up to 10 kilobases in length. The genes will be lassembled from individual oligonuclcotidcs synthesized in parallel on DNA arrays using the Maskless Array Synthesize_ (M.AS), which permits the production of custom DNA microarrays with 786,000 features in only 4 hours, Light-directed "safety-catch release chemistry will permit desired array components to be released from the surface in sets to form specific sub-assemblies, w/aich in turn will be assembled into the final long dsDNA product Biological error corredtion fiaethods, based upon the ability of the mismatch-binding protein MutS to recognize and remove mismatched duplexes, will_nsure high fidelity. All of these functions will be merged into an integrated system - the Automated Gene Synthesi_r (AGS) The ability to make complete genes on demand, inexpensively, and with rapid turn around time, has revolutionary implications for a wide range of biological and medical research The UW Bioengineering Partnership is made up of an exceptional interdisciplinary team of researchers with expertise in the areas of engineering, chemistry, inforrnatics, and molecular biology and cellular biology Team members have a proven track record i_t developing cutting edge tools for biological research. The inclusion of NimbleGen Systems on _e team, as an indusl_rial partner, brings additional resources in chemistry and MAS technology to accelerate development In addition to the primary focus of the group upon total gene synthesis, this bioengineering effort will also apply the massively p_rallel control of light to fuel basic research in the areas of combinatorial chemistry of small moldculcs and the detection of surface interactions Biological relevance of the technologies will be ensured by means of three specific applications: the rapid low cost production of DNA for gene targeting in stem cells (all five of the cell lines to be used in this proposal (HI, H7, H9, H13, and H14) are all listed on the NII-I registry), high-throughput protein synthesis, and high d_nsity on-chip SNP detection using sm'face invasive cleavage reactions "ERFORMANCESITE(S) (organizationc,/ty,state) Madison, Wl 53706 KEY PERSONNEL See instructions Start with Principal Investigator L}st all Name Cerrina, Francesco Beebe, David J Belshaw, PAter Kaysen, Ja_es H PatelJ Madl_usudan Richmond, Kathryn Shortfeed, Michael R Smith, Lloyd M Sussman, Michael R. Thomson, James A Zwaka. Thomas UseoontlnuaUopnagesasneededtoprovidetherequiredInformatiointheformatshownbelow other key personnel In,alphabetical order, last name first. Orgafiization Roleon Project UW-Madison Principal investigator Co-investigator UW]Madison Co-Investigator UW, Madison UW-Madison Senior Scientist UW-Madison Research Associate UW-Madison Assistant Scientist UW-Madison Assistant Scientist UW-Madison Co-Investigator UW-Madison Co-Investigator UW-Madison Co-Investigator UW-Madison Scientist, Project Leader Dlsc!n___ure Permlssfon SfafemenL Applicable to SBIR/STI'R Only_ Sea instructions [] Yes [] No PHS398(Rev 05/0t) I -_ Page2-- Number pages conseoutively at the bottom throughout FormPage Ihe application Do not use suffixes su[unreadable]has 2a, 2b ...... 9_:8g [unreadable]ggE-Tg-[unreadable]DO Principal Investigator/Program Director (Last, First, Middle): Cerrina, Francesco The name of the principal investigator/program director must be provided at the top of each printed page and each continuation page. RESEARCH GRANT TABLE OF CONTENTS Page Numbers Face Page ......................................................................................................................................................... 1 Description,